Dobrikova Elena Y, Grisham Rachel N, Kaiser Constanze, Lin Jennifer, Gromeier Matthias
Department of Molecular Genetics & Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
J Virol. 2006 Apr;80(7):3310-21. doi: 10.1128/JVI.80.7.3310-3321.2006.
Enteroviruses (EVs) overcome their host cells by usurping the translation machinery to benefit viral gene expression. This is accomplished through alternative translation initiation in a cap-independent manner at the viral internal ribosomal entry site (IRES). We have investigated the role of cis- and trans-acting viral factors in EV IRES translation in living cells. We observed that considerable portions of the viral genome, including the 5'-proximal open reading frame and the 3' untranslated region, contribute to stimulation of IRES-mediated translation. With the IRES in proper context, translation via internal initiation in uninfected cells is as efficient as at capped messages with short, unstructured 5' untranslated regions. IRES function is enhanced in cells infected with the EV coxsackievirus B3, but the related poliovirus has no significant stimulatory activity. This differential is due to the inherent properties of their 2A protease and is not coupled to 2A-mediated proteolytic degradation of the eukaryotic initiation factor 4G. Our results suggest that the efficiency of alternative translation initiation at EV IRESs depends on a properly configured template rather than on targeted alterations of the host cell translation machinery.
肠道病毒(EVs)通过篡夺翻译机制来实现对宿主细胞的感染,从而促进病毒基因表达。这是通过在病毒内部核糖体进入位点(IRES)以不依赖帽子结构的方式进行选择性翻译起始来完成的。我们研究了顺式和反式作用病毒因子在活细胞中EV IRES翻译中的作用。我们观察到病毒基因组的相当一部分,包括5'-近端开放阅读框和3'非翻译区,都有助于刺激IRES介导的翻译。在适当的背景下,未感染细胞中通过内部起始的翻译与具有短的、无结构的5'非翻译区的带帽信使RNA的翻译效率一样高。在感染肠道病毒柯萨奇病毒B3的细胞中IRES功能增强,但相关的脊髓灰质炎病毒没有显著的刺激活性。这种差异是由于它们2A蛋白酶的固有特性,并且与2A介导的真核起始因子4G的蛋白水解降解无关。我们的结果表明,EV IRESs处选择性翻译起始的效率取决于一个正确配置的模板,而不是宿主细胞翻译机制的靶向改变。